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Rapid, Scalable Assembly and Loading of Bioactive Proteins and Immunostimulants into Diverse Synthetic Nanocarriers Via Flash Nanoprecipitation

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Pneumatically controlled microfluidic synthesis of polymeric nanoparticles for mRNA delivery.

Yunshan Liu¹, Guanyou Lin¹, Matthew Michael James¹

¹Department of Materials Science and Engineering, University of Washington, 302L Roberts Hall, Seattle, Washington 98195, USA. mzhang@uw.edu.

|March 31, 2026

Summary

This study introduces a microfluidic platform for high-throughput synthesis of uniform polymeric nanoparticles (PNPs) for mRNA delivery. The technology offers a scalable and cost-effective method for producing advanced nucleic acid therapeutics.

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Area of Science:

Biotechnology and Nanotechnology
Materials Science and Engineering

Background:

Conventional bulk synthesis of polymeric nanoparticles (PNPs) for gene delivery faces challenges in reproducibility, scalability, and control over physicochemical properties.
Efficient gene delivery necessitates nanoscale carriers with precise characteristics, which are often not achievable with traditional manufacturing methods.

Purpose of the Study:

To develop a pneumatically driven microfluidic platform for precise, tunable, and high-throughput synthesis of polymeric nanoparticles for messenger RNA (mRNA) delivery.
To demonstrate the platform's capability in producing uniform and predictable nanoparticles with enhanced properties for therapeutic applications.

Main Methods:

A microfluidic device integrating a central air-driven channel with side reagent channels was designed to generate segmented gas-liquid flow.
Airflow modulation was employed to control nanoparticle formation, ensuring precise size control and enhanced mixing while preventing channel clogging.
The platform was used to synthesize PH-PEI-mRNA-heparin (PPH) nanoparticles.

Main Results:

The microfluidic platform produced PH-PEI-mRNA-heparin (PPH) nanoparticles with predictable, uniform sizes and extended chip lifespan.
Synthesized nanoparticles demonstrated low cytotoxicity, stable surface charge, efficient cellular uptake, and robust endosomal escape.
Transfection efficiency of the produced nanoparticles was comparable to or exceeded that of commercial lipid-based agents.

Conclusions:

The pneumatically driven microfluidic platform provides a scalable, reproducible, and cost-effective strategy for manufacturing polymeric mRNA nanocomplexes.
This versatile platform represents a significant advancement in the microfluidic production of nucleic acid therapeutics, offering improved control and efficiency.